Below is a gel of RT-PCR reactions similar to the ones you performed in exercise
ID: 162784 • Letter: B
Question
Below is a gel of RT-PCR reactions similar to the ones you performed in exercise 5.
a) What is the purpose of the DNAse treatment?
b) What is the purpose of the PCR reaction using RNA NOT treated with either DNAse or RT?
c) What were the sizes of the RT dependent and RT independent PCR products?
d) What does the difference in size between the PCR products tell you about the gene from which this RNA is derived from?
PLEASE EXPLAIN I WANT TO UNDERSTAND
M 1 2 3 4 5 Reaction Template PCR-1 R D (Step PCR-2 DR (step PCR-3 LRT-R (step III) PCR-4 IRT DR (step III PCR-5 east genomic DNAExplanation / Answer
In RT-PCR protocols, total RNA has been extracted from different cells, each cell has DNA and RNA, but we need only highly purified DNA free RNA. Thus, the RT-PCR protocol includes DNAase treatment to remove contamination with DNA in large RNA samples.
RNA used in RT-PCR quantitatively to detect the RNA expression through complementary DNA transcripts.
RT-independent PCR products agarose gel band resolution is poor but RT-dependent PCR bands has good resolution and give the accurate results
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